LOCUS       sequence-262889-        4155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4155)
  AUTHORS   Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, 
            Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, 
            Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov
            V, Wolf G, Crawford M, Everley R, ... Steppan CM
  TITLE     The RESOLUTE consortium: unlocking SLC transporters for drug 
            discovery.
  JOURNAL   Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 
            10.1038/d41573-020-00056-6.
  PUBMED    32265506
REFERENCE   2  (bases 1 to 4155)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nat Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7"; 
            pages: "429-430"
COMMENT     Sequence Label: pDONR221_SLC7A8
FEATURES             Location/Qualifiers
     source          1..4155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2280..2379)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2396..2415)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2397..2415)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2420..2436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2420..2436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2538..2557)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2549..3358
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2616..2635)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3505..4093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3994..4013
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggagg agggcgcccg gcacagaaac aataccgaga agaagcaccc
      721 aggcggcggc gagtccgacg cctctccaga ggcaggatct ggaggaggag gagtggccct
      781 gaagaaggag atcggcctgg tgagcgcctg cggcatcatc gtgggcaaca tcatcggctc
      841 cggcatcttc gtgtctccca agggcgtgct ggagaatgca ggaagcgtgg gactggccct
      901 gatcgtgtgg atcgtgaccg gctttatcac agtggtgggc gccctgtgct acgcagagct
      961 gggcgtgacc atcccaaaga gcggcggcga ctactcctat gtgaaggata tcttcggagg
     1021 actggcaggc tttctgaggc tgtggatcgc cgtgctggtc atctacccca ccaaccaggc
     1081 cgtgatcgcc ctgacattct ccaattatgt gctgcagcct ctgttcccaa cctgctttcc
     1141 ccctgagtct ggcctgagac tgctggccgc catctgcctg ctgctgctga catgggtgaa
     1201 ctgtagctcc gtgaggtggg caaccagggt gcaggatatc ttcacagcag gcaagctgct
     1261 ggccctggca ctgataatta taatgggcat tgtccagatc tgtaagggcg agtacttttg
     1321 gctggagcct aagaacgcct tcgagaattt tcaggagcca gacatcggac tggtggccct
     1381 ggccttcctg cagggctctt ttgcctacgg cggctggaac ttcctgaatt atgtgaccga
     1441 ggagctggtg gatccctata agaacctgcc tagggccatc tttatctcca tccctctggt
     1501 gaccttcgtg tacgtgtttg ccaacgtggc ctatgtgaca gccatgtctc cacaggagct
     1561 gctggccagc aatgccgtgg ccgtgacatt cggcgagaag ctgctgggcg tgatggcctg
     1621 gatcatgcct atctctgtgg ccctgagcac cttcggcggc gtgaatggca gcctgtttac
     1681 atctagccgg ctgttctttg caggagcaag agagggacac ctgccatccg tgctggccat
     1741 gatccacgtg aagcggtgca ccccaatccc cgccctgctg tttacctgta tctccacact
     1801 gctgatgctg gtgacctctg acatgtacac actgatcaac tatgtgggct tcatcaatta
     1861 cctgttttat ggcgtgacag tggccggcca gatcgtgctg aggtggaaga agccagatat
     1921 ccctcgccca atcaagatca acctgctgtt ccccatcatc tacctgctgt tctgggcctt
     1981 tctgctggtg tttagcctgt ggtccgagcc tgtggtgtgc ggaatcggac tggcaatcat
     2041 gctgaccgga gtgcccgtgt acttcctggg cgtgtattgg cagcacaagc ctaagtgctt
     2101 cagcgacttt atcgagctgc tgaccctggt gtcccagaag atgtgcgtgg tggtgtaccc
     2161 agaggtggag aggggatccg gcacagagga ggccaatgag gatatggagg agcagcagca
     2221 gcctatgtat cagcccaccc ctacaaagga caaggatgtg gccggccagc cacagcccga
     2281 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2341 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     2401 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2461 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2521 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2581 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2641 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttatt
     2701 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2761 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2821 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2881 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2941 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     3001 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     3061 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     3121 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3181 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3241 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3301 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3361 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3421 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3481 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3541 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3601 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3661 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3721 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3781 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3841 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3901 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3961 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4021 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4081 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4141 ttttgctcac atgtt
//
